Active Metabolite Norfluoxetine Research Articles

Toxicokinetics of the Antidepressant Fluoxetine and Its Active Metabolite Norfluoxetine in Caenorhabditis elegans and Their Comparative Potency.

The nematode Caenorhabditis elegans is a valuable model for ecotoxicological research, yet limited attention has been given to understanding how it absorbs, distributes, metabolizes, and excretes chemicals. This is crucial for C. elegans because the organism is known to have strong uptake barriers that are known to be susceptible to potential confounding effects of the presence of Escherichia coli as a food source. One frequently studied compound in C. elegans is the antidepressant fluoxetine, which has an active metabolite norfluoxetine. In this study, we evaluated the toxicokinetics and relative potency of norfluoxetine and fluoxetine in chemotaxis and activity tests. Toxicokinetics experiments were conducted with varying times, concentrations of fluoxetine, and in the absence or presence of E. coli, simulated with a one-compartment model. Our findings demonstrate that C. elegans can take up fluoxetine and convert it into norfluoxetine. Norfluoxetine proved slightly more potent and had a longer elimination half-life. The bioconcentration factor, uptake, and elimination rate constants depended on exposure levels, duration, and the presence of E. coli in the exposure medium. These findings expand our understanding of toxicokinetic modeling in C. elegans for different exposure scenarios, underlining the importance of considering norfluoxetine formation in exposure and bioactivity assessments of fluoxetine.

Influence of dissolved organic matter on the accumulation, metabolite production and multi-biological effects of environmentally relevant fluoxetine in crucian carp (Carassius auratus)

Fluoxetine is a widely prescribed antidepressant that has been frequently detected in aquatic environments and is associated with a series of neurological, behavioural and neuroendocrine disruptions in nontarget organisms. However, studies on its effects in fish under realistic environmental conditions are still limited. In this study, we determined the influences of an environmentally relevant concentration of fluoxetine (100 ng/L) on crucian carp (Carassius auratus) in the presence of dissolved organic matter (DOM). Endpoints that were assessed included accumulation of fluoxetine and metabolite formation as well as related biological responses involving neurotransmission and metabolic processes. Fluoxetine was significantly bioconcentrated in the fish brain and liver and largely transformed to the active metabolite norfluoxetine. Brain neurotransmission processes related to serotonin and choline and liver metabolic status were simultaneously altered. DOM added at 1 mg/L had no effect on the accumulation of fluoxetine or its metabolites in different tissues of the fish. However, at 10 mg/L DOM facilitated fluoxetine and norfluoxetine accumulation in the liver, brain, kidney, gill and bile tissues of the fish. The neuroendocrine-disrupting effects on fish caused by fluoxetine were also enhanced by the co-addition of DOM at 10 mg/L. Binding with fluoxetine and the inhibition of metabolic functions caused by DOM may be responsible for this increase in effects. These findings imply that at high concentrations DOM can increase the toxicity of environmentally relevant concentrations of fluoxetine to fish.

Simultaneous determination of fluoxetine, venlafaxine, vortioxetine and their active metabolites in human plasma by LC–MS/MS using one-step sample preparation procedure

The aim of antidepressant therapy is to induce remission and prevent relapses of major depressive disorder with minimum adverse effects during the treatment. Due to high variability in metabolism, therapeutic drug monitoring is recommended as a useful tool for individualisation of the therapy. For this purpose, we have developed simple and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for quantification of fluoxetine (FLX), venlafaxine (VEN), vortioxetine (VTX) and their active metabolites norfluoxetine (NFLX) and O-desmethylvenlafaxine (ODV). After one-step extraction procedure using OSTRO plate, analytes were separated by gradient elution on Acquity UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) column with runtime 4.2 min. The detection was done on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode with transitions at m/z 310.23 → 148.20 for FLX, m/z 296.23 → 134.20 for NFLX, m/z 278.31 → 121.13 for VEN, m/z 264.31 → 107.14 for ODV and m/z 299.19 → 150.05 for VTX using a positive electrospray ionisation interface. The method was successfully validated according to the European Medicine Agency guideline for the selectivity, linearity and lower limit of detection, precision and accuracy, matrix effect, extraction recovery, carryover, dilution integrity and stability over a concentration range of 1–300 ng/mL for FLX, NFLX, VEN, ODV and 0.2–100 ng/mL VTX. Extraction recovery for each analyte was > 80 %, and no significant matrix effects were observed. The developed method was employed for quantification of antidepressants in clinical samples from patients treated with either FLX, VEN, or VTX.

Serotonin and serotonin reuptake inhibitors alter placental aromatase

Serotonin reuptake inhibitors (SRIs) are currently the main molecules prescribed to pregnant women that suffer from depression. Placental cells are exposed to SRIs via maternal blood, and we have previously shown that SRIs alter feto-placental steroidogenesis in an in vitro co-culture model. More specifically, serotonin (5-HT) regulates the estrogen biosynthetic enzyme aromatase (cytochrome P450 19; CYP19), which is disrupted by fluoxetine and its active metabolite norfluoxetine in BeWo choriocarcinoma cells. Based on molecular simulations, the present study illustrates that the SRIs fluoxetine, norfluoxetine, paroxetine, sertraline, citalopram and venlafaxine exhibit binding affinity for the active-site pocket of CYP19, suggesting potential competitive inhibition. Using BeWo cells and primary villous trophoblast cells isolated from normal term placentas, we compared the effects of the SRIs on CYP19 activity. We observed that paroxetine and sertraline induce aromatase activity in BeWo cells, while venlafaxine, fluoxetine, paroxetine and sertraline decrease aromatase activity in primary villous trophoblast. The effects of paroxetine and sertraline in primary villous trophoblasts were observed at the lower doses tested. We also showed that 5-HT and the 5-HT2A receptor agonist 2,5-dimethoxy-4-iodoamphetamine (DOI) induced CYP19 activity. An increase in phosphorylation of serine and tyrosine and a decrease in threonine phosphorylation of CYP19 was also associated with DOI treatment. Our results contribute to better understanding how 5-HT and SRIs interact with CYP19 and may affect estrogen production. Moreover, this study suggests that alteration of placental 5-HT levels due to depression and/or SRI treatment during pregnancy may be associated with disruption of placental estrogen production.

Fluoxetine and its active metabolite norfluoxetine disrupt estrogen synthesis in a co-culture model of the feto-placental unit

The effects of fluoxetine, one of the most prescribed selective serotonin-reuptake inhibitors (SSRIs) during pregnancy, and its active metabolite norfluoxetine were studied on placental aromatase (CYP19) and feto-placental steroidogenesis. Fluoxetine did not alter estrogen secretion in co-culture of fetal-like adrenocortical (H295R) and trophoblast-like (BeWo) cells used as a model of the feto-placental unit, although it induced CYP19 activity, apparently mediated by the serotonin (5-HT)2A receptor/PKC signaling pathway. Norfluoxetine decreased estrogen secretion in the feto-placental co-culture and competitively inhibited catalytic CYP19 activity in BeWo cells. Decreased serotonin transporter (SERT) activity in the co-culture was comparable to 17β-estradiol treatment of BeWo cells. This work shows that the complex interaction of fluoxetine and norfluoxetine with placental estrogen production, involves 5-HT-dependent and -independent mechanisms. Considering the crucial role of estrogens during pregnancy, our results raise concern about the impact of SSRI treatment on placental function and fetal health.

Enantioselective biodegradation of fluoxetine by the bacterial strain Labrys portucalensis F11

Fluoxetine (FLX) is a chiral fluorinated pharmaceutical indicated mainly for the treatment of depression and is one of the most dispensed drugs in the world. There is clear evidence of environmental contamination with this drug and its active metabolite norfluoxetine (NFLX). In this study the enantioselective biodegradation of racemic FLX and of its enantiomers by Labrys portucalensis strain F11 was assessed. When 2μM of racemic FLX was supplemented as sole carbon source, complete removal of both enantiomers, with stoichiometric liberation of fluoride, was achieved in 30d. For racemic FLX concentration of 4 and 9μM, partial degradation of the enantiomers was obtained. In the presence of acetate as an additional carbon source, at 4, 9 and 21μM of racemic FLX and at 25μM of racemic FLX, (S)-FLX or (R)-FLX, complete degradation of the two enantiomers occurred. At higher concentrations of 45 and 89μM of racemic FLX, partial degradation was achieved. Preferential degradation of the (R)-enantiomer was observed in all experiments. To our knowledge, this is the first time that enantioselective biodegradation of FLX by a single bacterium is reported.

Fluoxetine dose and administration method differentially affect hippocampal plasticity in adult female rats.

Selective serotonin reuptake inhibitor medications are one of the most common treatments for mood disorders. In humans, these medications are taken orally, usually once per day. Unfortunately, administration of antidepressant medications in rodent models is often through injection, oral gavage, or minipump implant, all relatively stressful procedures. The aim of the present study was to investigate how administration of the commonly used SSRI, fluoxetine, via a wafer cookie, compares to fluoxetine administration using an osmotic minipump, with regards to serum drug levels and hippocampal plasticity. For this experiment, adult female Sprague-Dawley rats were divided over the two administration methods: (1) cookie and (2) osmotic minipump and three fluoxetine treatment doses: 0, 5, or 10 mg/kg/day. Results show that a fluoxetine dose of 5 mg/kg/day, but not 10 mg/kg/day, results in comparable serum levels of fluoxetine and its active metabolite norfluoxetine between the two administration methods. Furthermore, minipump administration of fluoxetine resulted in higher levels of cell proliferation in the granule cell layer (GCL) at a 5 mg dose compared to a 10 mg dose. Synaptophysin expression in the GCL, but not CA3, was significantly lower after fluoxetine treatment, regardless of administration method. These data suggest that the administration method and dose of fluoxetine can differentially affect hippocampal plasticity in the adult female rat.

Pharmacokinetics of Fluoxetine in Rhesus Macaques following Multiple Routes of Administration

Background/Aims: Fluoxetine (Prozac) is a selective serotonin reuptake inhibitor currently used to treat depression and mood disorders. It has been widely studied clinically and preclinically, yet there is limited knowledge of its pharmacokinetics in nonhuman primates. Methods: The present study characterized the pharmacokinetics of fluoxetine and its active metabolite norfluoxetine in rhesus macaques following both acute (1, 3, 5.6 and 10 mg/kg) and chronic doses (5.6 and 10 mg/kg/day) via different routes of administration (intravenous, subcutaneous, intramuscular, and oral). Blood samples were collected at multiple time points following administration and analyzed using mass spectrometry. Results: Fluoxetine had a half-life of 11–16 h and norfluoxetine had a half-life of 21–29 h. Potentially functionally significant serum concentrations of norfluoxetine were present at 24 h even after a single administration of fluoxetine. Similar to observations in humans under steady state conditions, norfluoxetine accounted for the greater percentage of active drug in the blood stream. Conclusion: A daily dose of 10 mg/kg administered orally maintained serum concentrations in the human clinical range over the course of 6 weeks. Given the long half-lives of fluoxetine and norfluoxetine observed in this study, precautions should be taken when designing preclinical studies to prevent accumulation of drug serum concentrations.

Pharmaceuticals as Neuroendocrine Disruptors: Lessons Learned from Fish on Prozac

Pharmaceuticals are increasingly detected in a variety of aquatic systems. One of the most prevalent environmental pharmaceuticals in North America and Europe is the antidepressant fluoxetine, a selective serotonin reuptake inhibitor (SSRI) and the active ingredient of Prozac. Usually detected in the range below 1 μg/L, fluoxetine and its active metabolite norfluoxetine are found to bioaccumulate in wild-caught fish, particularly in the brain. This has raised concerns over potential disruptive effects of neuroendocrine function in teleost fish, because of the known role of serotonin (5-HT) in the modulation of diverse physiological processes such as reproduction, food intake and growth, stress and multiple behaviors. This review describes the evolutionary conservation of the 5-HT transporter (the therapeutic target of SSRIs) and reviews the disruptive effects of fluoxetine on several physiological endpoints, including involvement of neuroendocrine mechanisms. Studies on the goldfish, Carassius auratus, whose neuroendocrine regulation of reproduction and food intake are well characterized, are described and represent a reliable model to study neuroendocrine disruption. In addition, fish studies investigating the effects of fluoxetine, not only on reproduction and food intake, but also on stress and behavior, are discussed to complement the emerging picture of neuroendocrine disruption of physiological systems in fish exposed to fluoxetine. Environmental relevance and key lessons learned from the effects of the antidepressant fluoxetine on fish are highlighted and may be helpful in designing targeted approaches for future risk assessments of pharmaceuticals disrupting the neuroendocrine system in general.

Trace analysis of fluoxetine and its metabolite norfluoxetine. Part I: Development of a chiral liquid chromatography–tandem mass spectrometry method for wastewater samples

An enantioselective method for the determination of fluoxetine (a selective serotonin reuptake inhibitor) and its pharmacologically active metabolite norfluoxetine has been developed for raw and treated wastewater samples. The stable isotope-labeled fluoxetine and norfluoxetine were used in an extended way for extraction recovery calculations at trace level concentrations in wastewater. Wastewater samples were enriched by solid phase extraction (SPE) with Evolute CX-50 extraction cartridges. The obtained extraction recoveries ranged between 65 and 82% in raw and treated wastewater at a trace level concentration of 50 pM (15–16 ng L −1). The target compounds were identified by the use of chiral liquid chromatography tandem mass spectrometry (LC–MS/MS) in selected reaction monitoring (SRM) mode. The enantiomers were successfully resolved on a chiral α 1-acid glycoprotein column (chiral AGP) with acetonitrile and 10 mM ammonium acetate buffer at pH 4.4 (3/97, v/v) as the mobile phase. The effects of pH, amount of organic modifier and buffer concentration in the mobile phase were investigated on the enantiomeric resolution ( R s) of the target compounds. Enantiomeric R s-values above 2.0 (1.03 RSD%, n = 3) were achieved for the enantiomers of fluoxetine and norfluoxetine in all mobile phases investigated. The method was validated by assessing parameters such as cross-contamination and carryover during SPE and during LC analysis. Cross-talk effects were examined during the detection of the analytes in SRM mode. In addition, the isotopic purity of fluoxetine-d 5 and norfluoxetine-d 5 were assessed to exclude the possibility of self-contamination. The interassay precision of the chromatographic separation was excellent, with relative standard deviations (RSD) equal to or lower than 0.56 and 0.81% in raw and treated wastewaters, respectively. The method detection and quantification limits (respectively, MDL and MQL) were determined by the use of fluoxetine-d 5 and norfluoxetine-d 5. The MQL for the single enantiomers ranged from 12 to 14 pM (3.6–4.3 ng L −1) in raw wastewater and from 3 to 4 pM (0.9–1 ng L −1) in treated wastewater. The developed method has been employed for the quantification of ( R)-fluoxetine, ( S)-fluoxetine and the enantiomers of norfluoxetine in raw and treated wastewater samples to be presented in Part II of this study.

Simultaneous determination of fluoxetine and its major active metabolite norfluoxetine in human plasma by LC‐MS/MS using supported liquid extraction

A rapid, sensitive and selective bioanalytical method was developed for the simultaneous determination of fluoxetine and its primary metabolite norfluoxetine in human plasma. Sample preparation was based on supported liquid extraction (SLE) using methyl tert-butyl ether to extract the analytes from human plasma. Chromatography was performed on a Synergi 4 μ polar-RP column using a fast gradient. The ionization was optimized using ESI (+) and selectivity was achieved by tandem mass spectrometric analysis using MRM functions, m/z 310 → 44 for fluoxetine, m/z 296 → 134 for norfluoxetine and m/z 315 → 44 for fluoxetine-d5 (internal standard). The method is linear over the range of 0.05-20 ng/mL (using a human plasma sample volume of 0.1 mL) with a coefficient determination of greater than 0.999. The method is accurate and precise with intra-batch and inter-batch accuracy (%bias) of < ± 15% and precision (%CV) of <15% for both analytes. A run time of 4 min means a high throughput of samples can be achieved. To our knowledge, this method appears to be the most sensitive one reported so far for the quantitation of fluoxetine and norfluoxetine and can be used for routine therapeutic drug monitoring or pharmacokinetic studies.

Waterborne fluoxetine disrupts the reproductive axis in sexually mature male goldfish, Carassius auratus

Fluoxetine (FLX) is a pharmaceutical acting as a selective serotonin reuptake inhibitor and is used to treat depression in humans. Fluoxetine and the major active metabolite norfluoxetine (NFLX) are released to aquatic systems via sewage-treatment effluents. They have been found to bioconcentrate in wild fish, raising concerns over potential endocrine disrupting effects. The objective of this study was to determine effects of waterborne FLX, including environmental concentrations, on the reproductive axis in sexually mature male goldfish. We initially cloned the goldfish serotonin transporter to investigate tissue and temporal expression of the serotonin transporter, the FLX target, in order to determine target tissues and sensitive exposure windows. Sexually mature male goldfish, which showed the highest levels of serotonin transporter expression in the neuroendocrine brain, were exposed to FLX at 0.54 μg/L and 54 μg/L in a 14-d exposure before receiving vehicle or sex pheromone stimulus consisting of either 4.3 nM 17,20β-dihydroxy-4-pregnene-3-one (17,20P) or 3 nM prostaglandin F 2α (PGF 2α). Reproductive endpoints assessed included gonadosomatic index, milt volume, and blood levels of the sex steroids testosterone and estradiol. Neuroendocrine function was investigated by measuring blood levels of luteinizing hormone, growth hormone, pituitary gene expression of luteinizing hormone, growth hormone and follicle-stimulating hormone and neuroendocrine brain expression of isotocin and vasotocin. To investigate changes at the gonadal level of the reproductive axis, testicular gene expression of the gonadotropin receptors, both the luteinizing hormone receptor and the follicle-stimulating hormone receptor, were measured as well as expression of the growth hormone receptor. To investigate potential impacts on spermatogenesis, testicular gene expression of the spermatogenesis marker vasa was measured and histological samples of testis were analyzed qualitatively. Estrogen indices were measured by expression and activity analysis of gonadal aromatase, as well as liver expression analysis of the estrogenic marker, esr1. After 14d, basal milt volume significantly decreased at 54 μg/L FLX while pheromone-stimulated milt volume decreased at 0.54 μg/L and 54 μg/L FLX. Fluoxetine (54 μg/L) inhibited both basal and pheromone-stimulated testosterone levels. Significant concentration-dependent reductions in follicle-stimulating hormone and isotocin expression were observed with FLX in the 17,20P- and PGF 2α-stimulated groups, respectively. Estradiol levels and expression of esr1 concentration-dependently increased with FLX. This study demonstrates that FLX disrupts reproductive physiology of male fish at environmentally relevant concentrations, and potential mechanisms are discussed.

GC-MS analysis of fluoxetine and its active metabolite norfluoxetine in human urine

ABSTRACT: A gas chromatographic-mass spectrometric (GC-MS) method was developed for detection of fluoxetine and its active metabolite norfluoxetine in urine. Liquid and solid phase extraction were applied to urine samples using maprotiline as an internal standard (IS). The GC-MS analysis were carried out using HP-5MS capillary column. The linearity ranges of the method were 5-75 ng mL-1 for fluoxetine and 6-125 ng mL-1 for norfluoxetine by solid phase extraction (SPE), and 10-80 ng mL-1 for fluoxetine and also norfluoxetine by liquid- liquid extraction (LLE). Also the range of detection limits were between 1-10 ng mL-1, the range of quantification limits were between 5-10 ng mL-1 for fluoxetine and norfluoxetine by both SPE and LLE. The range of recoveries were between 87 -109 % by both SPE and LLE for analytes. The developed method allowed clinical and toxicological analysis of fluoxetine and norfluoxetine in urine samples. KEY WORDS: Fluoxetine; Norfluoxetine; Gas Chromatography-Mass Spectrometry; Liquid- Liquid Extraction; Solid-Phase Extraction.

GC-MS analysis of fluoxetine and its active metabolite norfluoxetine in human urine

GC-MS analysis of fluoxetine and its active metabolite norfluoxetine in human urine

Physiological Modes of Action of Fluoxetine and its Human Metabolites in Algae

Fluoxetine, the active ingredient of many antidepressants, was identified as specifically toxic toward algae in a quantitative structure-activity relationship (QSAR) analysis with literature data for algae, daphnia, and fish. The goal of this study was to elucidate the mode of action in algae and to evaluate the toxicity of the major human metabolites of fluoxetine using two different algae tests. The time dependence and sensitivity of thedifferenteffectendpointsyield information on the physiological mode of action. Baseline toxicity was predicted with QSARs based on measured liposome-water partition coefficients. The ratio of predicted baseline toxicity to experimental toxicity (toxic ratio TR) gives information on the intrinsic potency (extent of specificity of effect). The metabolite p-trifluoromethylphenol was classified to act as baseline toxicant Fluoxetine (TR 60-150) and its pharmacologically active metabolite norfluoxetine (TR 10-80) exhibited specific toxicity. By comparison with reference compounds we conclude that fluoxetine and norfluoxetine have an effect on the energy budget of algal cells since the time pattern of these two compounds is most similar to that observed for norflurazon, but they act less specifically as indicated by lower TR values and the similarity of the effect pattern to baseline toxicants. The mixture toxicity of fluoxetine and its human metabolites norfluoxetine and p-TFMP can be predicted using the model of concentration addition for practical purposes of risk assessment despite small deviations from this model for the specific endpoints like PSII inhibition because the integrative endpoints like growth rate and reproduction in all cases gave agreement with the predictions for concentration addition.

Norfluoxetine Induces Spawning and Parturition in Estuarine and Freshwater Bivalves

Fluoxetine, a commonly prescribed antidepressant (Prozac), has been detected in sewage effluent. Its active metabolite norfluoxetine is more potent and has been detected in sewage influent and in fish tissues. We tested the effects of norfluoxetine on spawning and parturition in bivalves. Norfluoxetine induced significant spawning in zebra mussels and dark false mussels at concentrations as low as 5 microM. Norfluoxetine induced significant parturition in fingernail clams at 10 microM. Fluoxetine also induced spawning in dark false mussels at concentrations as low as 100 nM. Implications for environmental impacts of norfluoxetine and fluoxetine on native and exotic bivalves are discussed.

Determination of fluoxetine and its N-desmethyl metabolite in human plasma by high-performance liquid chromatography

A rapid and sensitive high-performance liquid chromatographic method has been developed for the simultaneous determination of the antidepressant fluoxetine and its active metabolite norfluoxetine in human plasma using paroxetine as internal standard. After liquid–liquid extraction, the compounds were separated on a C18 column using as mobile phase acetonitrile and 40 mM potassium dihydrogen phosphate buffer (pH 2.3) in the ratio 31:69 (v/v). The quantification of fluoxetine and norfluoxetine was made by fluorescence detection at Ex/Em 230/312 nm. The assay for each analyte was linear over the ranges 1–39 and 0.9–36 ng/ml, respectively. For both compounds intra- and inter-day accuracy and precision ranged between −7.9–12.4 and 0.7–14.7%, respectively. The method was applied to the analysis of plasma samples obtained from healthy subjects treated with one single oral dose of 40 mg fluoxetine.

Determination of fluoxetine and its major active metabolite norfluoxetine in human plasma by liquid chromatography-tandem mass spectrometry

A sensitive and specific bioanalytical method for the determination of fluoxetine and norfluoxetine in human plasma has been developed. Automated solid phase extraction on Oasis HLB cartridges was used to extract the analytes from human plasma. Analysis was by reverse phase liquid chromatography on a Xterra MS C18 column using a fast gradient. Fluoxetine, norfluoxetine and fluvoxamine (internal standard) were ionised using the Turbolonspray interface operating in positive ion mode. Detection was via multiple reaction monitoring (MRM) of the characteristic ion dissociation transitionsm/z 310.3→44.1, 296.2→134.3 and 319.2→71.1 for fluoxetine, norfluoxetine and fluvoxamine respectively. The method is linear over the range 0.5–50 ng mL−1 (using a sample volume of 0.5 mL). The method is accurate and precise with intra-batch and inter-batch precision (%CV) of<15% and accuracy (%RE) of <+-15% for both analytes. A run time of 4 minutes means a high throughput of samples can be achieved. The method has been be used to support a clinical study.

Sensitive liquid chromatographic–tandem mass spectrometric method for the determination of fluoxetine and its primary active metabolite norfluoxetine in human plasma

A sensitive method for the simultaneous determination of fluoxetine and its major active metabolite norfluoxetine in plasma was developed, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The samples were extracted from alkalised plasma with hexane–isoamyl alcohol (98:2, v/v) followed by back-extraction into formic acid (2%). Chromatography was performed on a Phenomenex ® Luna C 18 (2) 5 μm, 150×2 mm column with a mobile phase consisting of acetonitrile–0.02% formic acid (340:660, v/v) at a flow-rate of 0.35 ml/min. Detection was achieved by a Perkin-Elmer Sciex API 2000 mass spectrometer (LC–MS–MS) set at unit resolution in the multiple reaction monitoring mode. TurboIonSpray ionisation was used for ion production. The mean recoveries for fluoxetine and norfluoxetine were 98 and 97%, respectively, with a lower limit of quantification set at 0.15 ng/ml for the analyte and its metabolite. This assay method makes use of the increased sensitivity and selectivity of mass spectrometric (MS–MS) detection to allow for a more rapid (extraction and chromatography) and sensitive method for the simultaneous determination of fluoxetine and norfluoxetine in human plasma than has previously been described.

Pharmacokinetic drug interaction potential of selective serotonin reuptake inhibitors.

Obsessive-compulsive disorder (OCD) is a chronic disorder requiring long-term treatment. The pharmacological management of the disorder, therefore, requires the use of agents which, in addition to being efficacious and well tolerated, are unlikely to cause pharmacokinetic drug-drug interactions with concomitantly administered medication which the patient is receiving or may receive in the future. The selective serotonin reuptake inhibitors (SSRIs) have similar pharmacodynamic profiles but their pharmacokinetic profiles are very different. Perhaps the most substantial pharmacokinetic difference among these drugs is in their potential for drug-drug interactions via the inhibition of cytochrome P450 (CYP) isoenzymes. This review provides comprehensive background information on these enzyme systems and discusses their significance with respect to the optimal care of patients. Fluoxetine is a substantial inhibitor of CYP2D6, has mild effects on CYP3A3/4, and may also have effects on CYP2C9/10 and CYP2C19. Effects on drugs metabolized by these enzymes can persist for many weeks after fluxoetine discontinuation due to the long half-life of fluoxetine and its active metabolite norfluoxetine. Fluvoxamine is a substantial inhibitor of CYP1A2 and CYP2C19, and a moderate inhibitor of CYP3A3/4. Paroxetine is a substantial inhibitor of CYP2D6. In contrast, sertraline and citalopram are mild inhibitors of CYP2D6 at their usually effective doses and are not known to produce clinically meaningful inhibition of any other isoenzymes. However, citalopram has not been well studied against all of these enzymes, especially in vivo. An increased risk of pharmacokinetic drug interactions is the immediate clinical consequence of the inhibitory effects of these drugs on CYP isoenzymes. However, with the emphasis on long-term treatment, particularly in chronic conditions such as OCD, it will also be important to determine the long-term clinical consequences of substantially inhibiting specific CYP isoenzymes with those SSRIs which have these effects. Thus knowledge of the substrates and inhibitors of CYP isoenzymes may help clinicians to anticipate and avoid pharmacokinetic drug interactions and may better define rational prescribing practices.